Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding process.ZmPLA1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,ex...Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding process.ZmPLA1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,exhibiting a synergistic effect.However,it is unknown whether knock out of ZmDMP orthologs can stimulate HI in rice.In this study,a ZmPLA1 ortholog(OsPLA1)and two ZmDMP orthologs(OsDMP3 and OsDMP6)were identified in rice.All three genes encode plasma membrane-localized proteins and were highly expressed in mature anthers.Knockout of OsPLA1 in both Minghui 63 and Nipponbare resulted in reduced seed setting rate(SSR)and caused HI.The osdmp3,osdmp6 and the double mutant failed to trigger HI independently,nor increased the haploid induction rate(HIR)when combined with ospla1.Repeated pollinations operations of QX654A with the ospla1 mutant significantly improve SSR,while reducing HIR.RNA-seq profiling of mature ospla1 mutant anthers indicated that a large number of differentially expressed genes(DEGs)were enriched in redox homeostasis and lipid metabolic GO terms,plant hormone signal transduction,and MAPK signaling pathways.These findings provide important insights towards construction of an efficient DH breeding technology and study of the molecular mechanism of HI in rice.展开更多
The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome edit...The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome editing(GE).Recently,these techniques were combined into a GE trait delivery system called HI-Edit(Haploid Inducer-Edit).In HI-Edit,the pollen of a haploid inducer line is reprogrammed to deliver GE traits to any variety,obviating recurrent selection.For HI-Edit to operate at scale,an efficient transformable HI line is needed,but most maize varieties are recalcitrant to transformation,and haploid inducers are especially difficult to transform given their aberrant reproductive behaviors.Leveraging marker assisted selection and a three-tiered testing scheme,we report the development of new Iodent and Stiff Stalk maize germplasm that are transformable,have high haploid induction rates,and exhibit a robust,genetically-dominant anthocyanin native trait that may be used for rapid haploid identification.We show that transformation of these elite‘‘HI-Edit”lines is enhanced using the BABYBOOM and WUSCHEL morphogenetic factors.Finally,we evaluate the HI-Edit performance of one of the lines against both Stiff Stalk and non-Stiff Stalk testers.The strategy and results of this study should facilitate the development of commercially scalable HI-Edit systems in diverse crops.展开更多
Doubled haploid(DH)plants have been widely used for breeding and biological research in crops.Pop ulus spp.have been used as model woody plant species for biological research.However,the induction of DH poplar plants ...Doubled haploid(DH)plants have been widely used for breeding and biological research in crops.Pop ulus spp.have been used as model woody plant species for biological research.However,the induction of DH poplar plants is onerous,and limited biological or breeding work has been carried out on DH individuals or populations.In this study,we provide an effective protocol for poplar haploid induction based on an anther culture method.A total of 96 whole DH plant lines were obtained using an F1hybrid of Populus simonii×P.nigra as a donor tree.The phenotypes of the DH population showed exceptionally high variance when compared to those of half-sib progeny of the donor tree.Each DH line displayed distinct features compared to those of the other DH lines or the donor tree.Additionally,some excellent homozygous lines have the potential to be model plants in genetic and breeding studies.展开更多
BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromi...BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens.AIM To derive human haploid neural stem cells(haNSCs)to carry out lineage-specific screens.METHODS Human haNSCs were differentiated from human extended haESCs with the help of Y27632(ROCK signaling pathway inhibitor)and a series of cytokines to reduce diploidization.Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency.Global gene expression analysis was conducted to compare haNSCs with diploid NSCs and haESCs.Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs.Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools.RESULTS Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies,a higher efficiency in neural differentiation,a higher cell survival ratio and higher stability in haploidy maintenance.These characteristics effectively facilitated the derivation of human haNSCs.These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro.After PiggyBac transfection,there were multiple insertion sites in the human haNSCs’genome,and the insertion sites were evenly spread across all chromosomes.In addition,after the cells were treated with manganese,we were able to generate a list of manganese-induced toxicity genes,demonstrating their utility as genetic screening tools.CONCLUSION This is the first report of a generated human haploid somatic cell line with a complete genome,proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.展开更多
Doubled haploid(DH) technology is important in modern maize breeding. Haploid inducers determine the efficiency of both haploid induction and identification. It has taken decades to improve the efficiency,haploid indu...Doubled haploid(DH) technology is important in modern maize breeding. Haploid inducers determine the efficiency of both haploid induction and identification. It has taken decades to improve the efficiency,haploid induction rate(HIR), from the ~2% of the ancestor haploid inducer, stock6, to the ~10% of modern haploid inducers. Improvement of kernel oil content(KOC) would further enhance haploid identification efficiency. Using molecular marker-assisted selection, in combine with the number of haploids per ear as phenotypic criterion, we developed a new high-oil haploid inducer line, CHOI4, with a mean HIR of 15.8%and mean KOC of 11%. High KOC of CHOI4 can achieve a mean accuracy greater than 90% in identification of haploids of different backgrounds, with reduced false discovery rates and false negative rates in comparison with the previous high-oil haploid inducer line, CHOI3. Comparison of phenotypic selection strategies suggested that the number of haploids per ear can be used as a phenotyping criterion during haploid inducer line development. CHOI4 could further increase the efficiency of large-scale DH breeding programs with lower cost.展开更多
There is no spontaneous chromosome doubling in haploid plants produced by wheat × maize crossing. In order to obtain doubled haploid, two chromosome doubling methods were used. Results showed that: After adding c...There is no spontaneous chromosome doubling in haploid plants produced by wheat × maize crossing. In order to obtain doubled haploid, two chromosome doubling methods were used. Results showed that: After adding colchicine solution directly into a medium for young embryos that had been cultured 7 days,frequencies of embryo germination in colchicine concentrations of 50lng/L, 100mg/L and 200mg/L were 32.1%, 26.4% and 16.3%, respectively, and frequencies of chromosome doubling were 85.3%, 100% and 50.0%, respectively. But in the control without colchicine, the frequency of embryo germination was 67.4%and no seed was setting. As the time of colchicine treatment increased from 24 to 72 hours, the frequency of embryo germination was reduced, and 24 hours had better results. After soaking seeding crowns and roots with colchicine solution of 500mg/L, 750mg/L and 1 000mg/L for 5 hours, the frequencies of doubling were 89.6%, 76.0% and 73.3%, respectively. By soaking crowns and roots of strong seedings with 500mg/L colchicine solution, the frequency and efficiency of doubling were 98.2% and 93.2%, respectively.展开更多
Artificial gynogenesis is of great research value in fish genetics and breeding technology. However, existing studies did not explain the mechanism of some interesting phenomena. Severe developmental defects in gynoge...Artificial gynogenesis is of great research value in fish genetics and breeding technology. However, existing studies did not explain the mechanism of some interesting phenomena. Severe developmental defects in gynogenetic haploids can lead to death during hatching. After diploidization of chromosomes, gynogenetic diploids may dispense from the remarkable malformation and restore the viability, although the development time is longer and the survival rate is lower compared with normal diploids. The aim of this study was to reveal key mechanism in haploid syndrome of Japanese flounder, a commercially important marine teleost in East Asia. We measured genome-scale gene expression of flounder haploid, gynogenetic diploid and normal diploid embryos using RNA-Seq, constructed a module-centric co-expression network based on weighted correlation network analysis(WGCNA) and analyzed the biological functions of correlated modules. Module gene content analysis revealed that the formation of gynogenetic haploids was closely related to the abnormality of plasma proteins, and the up-regulation of p53 signaling pathway might rescue gynogenetic embryos from haploid syndrome via regulating cell cycle arrest, apoptosis and DNA repair. Moreover, normal diploid has more robust nervous system. This work provides novel insights into molecular mechanisms in haploid syndrome and the rescue process by gynogenetic diploidization.展开更多
A reliable,efficient anther culture system,the dominant technique for generating haploid plants in breeding programs,that can be used for generating transgenic poplar plants has been needed.In the present study,theref...A reliable,efficient anther culture system,the dominant technique for generating haploid plants in breeding programs,that can be used for generating transgenic poplar plants has been needed.In the present study,therefore,an anther culture system was developed using isolated mid-and late-uninucleate anthers of poplar(Populus simonii x P.nigra).From a combination of SSR and ploidy analyses,six double haploid and two haploid lines were characterized from 86 plants grown from 16 regenerated anther cultured lines.After 48 months of development,two plant lines from the regenerated plants maintained their haploid level in vitro for over 2 years.A number of haploid plants from the different lines weretransferred to soil.The leaves of these transplants were then used as explants for transformation with the APETALA1(AP1) gene using Agrobacterium tumefaciens.Overexpression of AP1 in haploid poplar induced early flowering with obvious petals when ectopically expressed.To our knowledge,this is the first report on changes in flowering time in AP1-trangenic poplar,which is important for elucidating the regulatory mechanism of tree flower development.展开更多
To reveal the sterility characteristics of Lycium barbarum haploids, cytological observations were made on the anthers of Ningqi No.1 and its haploids obtained from anther culture. The results showed that there were n...To reveal the sterility characteristics of Lycium barbarum haploids, cytological observations were made on the anthers of Ningqi No.1 and its haploids obtained from anther culture. The results showed that there were no significant differences in anther development between Ningqi No.1 and its haploids at the stage of pollen mother cell, and tetrads were formed successfully after the meiosis stage. The tetrads of Ningqi No.1 could release microspores. At the same time, the tapetal cells can provide nutrition for the development of the microspores, which eventually developed into mature pollen grains. Although the haploids could also release microspores at the tetrad stage, the tapetal cells degraded in advance, which made the released microspores unable to develop into mature pollen grains, resulting in pollen abortion of haploids.展开更多
Doubled haploid(DH)breeding technology,which relies on haploid genome doubling,is widely used in commercial maize breeding.Spontaneous haploid genome doubling(SHGD),a more simplified and straightforward method,is gain...Doubled haploid(DH)breeding technology,which relies on haploid genome doubling,is widely used in commercial maize breeding.Spontaneous haploid genome doubling(SHGD),a more simplified and straightforward method,is gaining popularity among maize breeders.However,the cytological mechanism of SHGD remains unclear.This study crossed inbred lines RL36 and RL7,which have differing SHGD abilities,with inducer line YHI-1 to obtain haploid kernels.The meiotic processes of pollen mother cells(PMCs)in the haploid plants were compared with diploid controls.The results suggested that three main pathways,the early doubling of haploid PMCs,the first meiotic metaphase chromosomal segregation distortion,and anomaly of the second meiosis,are responsible for SHGD.Furthermore,flow cytometry analysis of ploidy levels in leaves and PMCs from haploids and diploid controls revealed that somatic cell chromosome doubling and germ cell chromosome doubling are independent processes.These findings provide a foundation for further studies on the underlying mechanism of SHGD,aiding the application of DH technology in maize breeding practices.展开更多
This study determined the indirect selection index of doubled haploid(DH)rice using a multivariate analysis approach to select lines adaptive to salinity stress,comprising three experiments.The first experiment involv...This study determined the indirect selection index of doubled haploid(DH)rice using a multivariate analysis approach to select lines adaptive to salinity stress,comprising three experiments.The first experiment involved the selection of good agronomic characters in a field experiment conducted at an experimental station in Bogor,Indonesia.The second experiment involved salinity tolerance screening through hydroponic cultures using 0 and 120 mmol/L NaCl,conducted at a greenhouse in Bogor.The third experiment involved the validation of the indirect adaptability selection index(IASI)through a field experiment in Sukra(saline area).Field experiments followed a randomized complete block design(RCBD),whereas an RCBD nested factorial design was used for the greenhouse experiment.The first and second experiments used 56 DH lines and four check varieties with three replications.In the second experiment,Pokkali and IR29 varieties were also added as tolerant and sensitive checks of salinity,respectively.The third experiment used 28 selected DH lines,Inpari 29 and one sensitive DH line.The good agronomic index(GAI)was 0.465 yield+0.433 number of productive tillers+0.31 number of filled grains.This generated 24 DH rice lines with good agronomic traits.The salinity tolerance index(SaTI)was developed through the average of standardized salinity tolerance score and salinity selection index based on discriminant analysis.This generated 34 DH rice lines with good salinity stress tolerance.The IASI(IASI=GAI–SaTI)selected 28 DH rice lines adaptive to salinity stress and it was considered effective by Sukra validation.展开更多
The rye genome has shown potential for improvement of bread wheat,where wheat-rye substitutions and translocations have been and are frequently used in resistance breeding.Crosses belongs to different generations viz....The rye genome has shown potential for improvement of bread wheat,where wheat-rye substitutions and translocations have been and are frequently used in resistance breeding.Crosses belongs to different generations viz.,BC_(1)F_(1),BC_(1)F_(2),BC_(1)F_(3),BC_(1)F_(4)and BC_(2)F_(3)of triticale×wheat derived were used for different haploid induction parameters using Gogon grass(Imperata cylindrica)as a pollen source.The percentage of pseudo seed formation ranged from 34.55%for BC_(1)F_(2)to 63.77 for BC_(1)F_(1)crosses,the haploid embryo formation ranges from 9.43%for BC_(1)F_(1)to 30.2%for BC_(1)F_(2),the haploid plant generation ranges from 19.36%for BC_(1)F_(2)to 63.25%for BC_(1)F_(1).Four doubled haploids were developed from ITSN 105/58-VL 802-VL 802 of BC_(2)F_(3)underwent molecular cytogenetic analyses using the probes,viz.,rye genomic rDNA,pSc 119 and pAs1.FISH and GISH analysis revealed an IBL.1RS translocation and substitution of 5R chromosome instead of the 5D chromosomes in these doubled haploids.展开更多
Stability analysis of grain yield is an efficient tool for the selection of varieties adapted to fairly wide cultivation zone. A study aims to determine adaptability and stability for grain yield and agronomic perform...Stability analysis of grain yield is an efficient tool for the selection of varieties adapted to fairly wide cultivation zone. A study aims to determine adaptability and stability for grain yield and agronomic performance of 10 elite Doubled Haploid (DH) rice genotypes generated through another culture technique, along with local cultivar Umgar as a check under irrigated lowland conditions in three different environments in the central clay plains of the Sudan;during two cropping seasons 2019 and 2020. The trials were conducted in a randomized complete block design with three replications. Grain yield stability was studied, using the Additive Main effect and Multiplicative Interaction (AMMI) model. Bi-plots were developed following GGE bi-plot methodology over the six environments. The combined analysis of variance revealed significant to highly significant (P < 0.001) effects of genotypes, environments, and genotype by environment interaction. Moreover, the analysis of variance based on AMMI indicated significant genotypes, environments, and GE interaction with a total variation of 43.16%, 44.26% and 12.58% respectively. A biplot-AMMI analysis and yield stability index incorporating the AMMI stability value and yield in a single non-parametric index were used to identify the genotypes with the highest and stable yield. The overall mean for grain yield was 5.69 t/ha, with a range of 5.77 to 4.12 t/ha. The genotypes k150147, k150870, k150032, k150912, and k150307 out yielded the local check Umgar by 40%, 38%, 34%, 34% and 33% respectively. The most genotypes were mid-early maturities which were harvested at 107, 113, 108, 109 and 112 days after sowing, respectively. These five genotypes also showed grain yield stability along with their high mean yield performance according to the AMMI analysis and widely adaptable to the tested locations. Therefore, the five out yielding genotypes will be considered for cultivation under irrigated system condition in central clay in the Sudan.展开更多
The development of new rice varieties is highly dependent on genetic diversity in desirable agronomic traits. Therefore, this study aimed to identify potential genotypes having the characters of Korean varieties (Tong...The development of new rice varieties is highly dependent on genetic diversity in desirable agronomic traits. Therefore, this study aimed to identify potential genotypes having the characters of Korean varieties (Tongil-type) and japonica developed through doubled haploid (DH) technology to apply in our breeding materials. 35 elite DH lines derived from another culture of Korean and African rice along with two local checks were planted in a randomized complete block design with three replications during the two seasons of 2019 and 2020. All evaluated genotypes exhibited a wide and significant variation in the ten measured traits. The highest heritability related to high genetic advance was recorded for the number of tiller/plant, grain yield t/ha, number of filled grain per panicle, and thousand grain weights (g). Genotypic coefficient of variation and genetic advance were recorded for number of tiller/plant, number of filled grain per panicle, 1000 grain weight and grain yield t/ha in both seasons. Moreover, there was a highly significant and positive correlation of grain yield with number of filled grain per panicle (0.65), number of tiller/plant (0.64) and number of panicle per m<sup>2</sup> (0.54). Cluster analysis based on grain yield components trait grouped the 37 rice genotypes into four clusters. Cluster B was the largest and consisted of 13 genotypes. Finally, it could be concluded that, based on number of productive tillers, number of filled grain/panicle, number of panicle per m<sup>2</sup>, and grain yield, the lines KF170506, KF170509, KF170542, KF170530, KF170543, KF170500 and KF170510 were high potential for further selection for new type of irrigated rice. In addition, hybridization of these 7 high-yielding could be used to achieve higher heterosis among the genotypes. Furthermore, this evaluation could be useful in developing reliable selection indices for improving rice breeding programs.展开更多
Quantitative trait loci (QTLs) for plant height in wheat (Triticum aestivum L.) were studied using a set of 168 doubled haploid (DH) lines, which were derived from the cross Huapei 3/Yumai 57. A genetic linkage map wa...Quantitative trait loci (QTLs) for plant height in wheat (Triticum aestivum L.) were studied using a set of 168 doubled haploid (DH) lines, which were derived from the cross Huapei 3/Yumai 57. A genetic linkage map was constructed using 283 SSR and 22 EST-SSR markers. The DH population and the parents were evaluated for wheat plant height in 2005 and 2006 in Tai’an and 2006 in Suzhou. QTL analyses were performed using the software of QTLNetwork version 2.0 based on the mixed linear model. Four additive QTLs and five pairs of epistatic effects were detected, which were distributed on chromosomes 3A, 4B, 4D, 5A, 6A, 7B, and 7D. Among them, three additive QTLs and three pairs of epistatic QTLs showed QTL×environment interactions (QEs). Two major QTLs, Qph4B and Qph4D, which accounted for 14.51% and 20.22% of the phenotypic variation, were located similar to the reported locations of the dwarfing genes Rht1 and Rht2, respectively. The Qph3A-2 with additive effect was not reported in previous linkage mapping studies. The total QTL effects detected for the plant height explained 85.04% of the phenotypic variation, with additive effects 46.07%, epistatic effects 19.89%, and QEs 19.09%. The results showed that both additive effects and epistatic effects were important genetic bases of wheat plant height, which were subjected to environmental modifications, and caused dramatic changes in phenotypic effects. The information obtained in this study will be useful for manipulating the QTLs for wheat plant height by molecular marker-assisted selection (MAS).展开更多
A new doubled haploid (DH) rice population was established from a cross between WBPH-resistant japonica Chunjiang 06 (C J-06) and susceptible indica TN1. Sucking inhibitory and ovicidal resistance of the DH rice lines...A new doubled haploid (DH) rice population was established from a cross between WBPH-resistant japonica Chunjiang 06 (C J-06) and susceptible indica TN1. Sucking inhibitory and ovicidal resistance of the DH rice lines were evaluated on the basis of non-preference response of WBPH immigrants and honeydew excretion by WBPH females, and appearance of watery lesions in the necrotic discoloration of leaf sheaths ovipositied by WBPH,respectively. Both the major gene resistance to WBPH, sucking inhibitory and ovicidal resistance, showed 1 (resistant): 1 (susceptible) segregation ratio in the DH population. Relative density of WBPH populations and damage scores in the DH population indicated combined functions of both the major resistance genes as well as QTLs affecting the host plant response to WBPH infestations. Thus, the newly developed CJ-06/-I-N1 DH population could be a useful material to analyze major genes and QTLs for WBPH resistance in japonica rice.展开更多
In order to determine the potential for haploid induction via in vitro gynogenesis in tomato, the ovules and protoplasts of embryo sacs from the hybrids Zhongza 101 and Zhongza 105 were cultured. An efficient method o...In order to determine the potential for haploid induction via in vitro gynogenesis in tomato, the ovules and protoplasts of embryo sacs from the hybrids Zhongza 101 and Zhongza 105 were cultured. An efficient method of ovule isolation was established in this study. Using this method, 100-150 ovules could be isolated from one ovary. Isolated ovules were cultured on three induction media to induce gynogenesis in vitro. During culture, ovules were enlarged markedly, with opaque white color. When observed microscopically, there were cell divisions and cell clumps in embryo sacs. Subsequently, the cell clumps in embryo sacs ceased growth, likely because the integument grew faster than embryo sacs did and hindered the further development of embryo sacs. Therefore, subsequent callus morphogenesis might be originated from the integument. Thousands of calli from the two tomato varieties were obtained. Five diploid plants were regenerated after 15 months of subculturing. To eliminate the hindering effect of integument on embryo sac cells, the protoplasts of embryo sacs were prepared and cultured. After 48 hours of culture, the protoplasts of embryo sacs doubled in size and gradually formed clusters of cells. These results suggested that gynogenesis might be a potential way for haploid induction in tomato.展开更多
Sheath blight(SB) disease,caused by Rhizoctonia solani K(u|¨)hn,is one of the most serious diseases causing rice(Oryza sativa L.) yield loss worldwide.A doubled haploid(DH) population was constructed from a cross...Sheath blight(SB) disease,caused by Rhizoctonia solani K(u|¨)hn,is one of the most serious diseases causing rice(Oryza sativa L.) yield loss worldwide.A doubled haploid(DH) population was constructed from a cross between a japonica variety CJ06 and an indica variety TN1,and to analyze the quantitative trait loci(QTLs) for SB resistance under three different environments(environments 1-3).Two traits were recorded to evaluate the SB resistance,namely lesion height(LH) and disease rating(DR).Based on field evaluation of SB resistance and a genetic map constructed with 214 markers,a total of eight QTLs were identified for LH and eight QTLs for DR under three environments,respectively.The QTLs for LH were anchored on chromosomes 1,3,4,5,6,and 8,and explained 4.35-17.53%of the phenotypic variation.The SB resistance allele of qHNLH4 from TN1 decreased LH by 3.08 cm,and contributed to 17.53%of the variation at environment 1.The QTL for LH(qHZaLH8) detected on chromosome 8 in environment 2 explained 16.71%of the variation,and the resistance allele from CJ06 reduced LH by 4.4 cm.Eight QTLs for DR were identified on chromosomes 1,5,6,8,9,11,and 12 under three conditions with the explained variation from 2.0 to 11.27%.The QTL for DR(qHZaDR8),which explained variation of 11.27%,was located in the same interval as that of qHZaLH8,both QTLs were detected in environment 2.A total of six pairs of digenic epistatic loci for DR were detected in three conditions,but no epistatic locus was observed for LH.In addition,we detected 12 QTLs for plant height(PH) in three environments.None of the PH-QTLs were co-located with the SB-QTLs.The results facilitate our understanding of the genetic basis for SB resistance in rice.展开更多
Whitebacked planthopper (WBPH) -resistance in a japonica / indica doubled haploid (DH) rice population established from a cross between WBPH-resistant japonica Chunjiang 06 and susceptible indica TN1, was comparativel...Whitebacked planthopper (WBPH) -resistance in a japonica / indica doubled haploid (DH) rice population established from a cross between WBPH-resistant japonica Chunjiang 06 and susceptible indica TN1, was comparatively evaluated through a field experiment based on the WBPH immigrant density and standardized seedbox screening test (SSST). All the susceptible DH lines in the field experiment behaved accordingly in SSST. However, 35 of resistant 66 lines (53%) in the field, were categorized to susceptible groups in SSST. Likewise, there were no significant differences in WBPH immigrant densities among 70 DH lines that were highly resistant to susceptible in SSST. The results revealed that SSST could not evaluate properly WBPH resistance in the DH lines. Four QTLs for WBPH-resistance phenotyped by the immigrant density were detected on chromosomes 2, 3, 4, and 11. Of them, the QTL on chromosome 4 was the most effective (LOD 21.8, variance 78%). Five QTLs associated with seedling mortality were mapped on chromosomes 2, 3, 4, 5 and 6. In addition to the QTL (LOD 10.5, variance 68%) on chromosome 4, there was another major QTL (LOD 12.7, variance 71%) located on chromosome 5, which was SSST-specific but might be irrespective of the WBPH resistance traits.展开更多
基金This work was supported by the National Key Research and Development Program of China(2022YFD1200800)the China Agriculture Research System(CARS-02-05)+1 种基金Beijing Nova Program(2023067)Yunnan Province Science and Technology Department(202305AF150026).
文摘Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding process.ZmPLA1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,exhibiting a synergistic effect.However,it is unknown whether knock out of ZmDMP orthologs can stimulate HI in rice.In this study,a ZmPLA1 ortholog(OsPLA1)and two ZmDMP orthologs(OsDMP3 and OsDMP6)were identified in rice.All three genes encode plasma membrane-localized proteins and were highly expressed in mature anthers.Knockout of OsPLA1 in both Minghui 63 and Nipponbare resulted in reduced seed setting rate(SSR)and caused HI.The osdmp3,osdmp6 and the double mutant failed to trigger HI independently,nor increased the haploid induction rate(HIR)when combined with ospla1.Repeated pollinations operations of QX654A with the ospla1 mutant significantly improve SSR,while reducing HIR.RNA-seq profiling of mature ospla1 mutant anthers indicated that a large number of differentially expressed genes(DEGs)were enriched in redox homeostasis and lipid metabolic GO terms,plant hormone signal transduction,and MAPK signaling pathways.These findings provide important insights towards construction of an efficient DH breeding technology and study of the molecular mechanism of HI in rice.
文摘The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome editing(GE).Recently,these techniques were combined into a GE trait delivery system called HI-Edit(Haploid Inducer-Edit).In HI-Edit,the pollen of a haploid inducer line is reprogrammed to deliver GE traits to any variety,obviating recurrent selection.For HI-Edit to operate at scale,an efficient transformable HI line is needed,but most maize varieties are recalcitrant to transformation,and haploid inducers are especially difficult to transform given their aberrant reproductive behaviors.Leveraging marker assisted selection and a three-tiered testing scheme,we report the development of new Iodent and Stiff Stalk maize germplasm that are transformable,have high haploid induction rates,and exhibit a robust,genetically-dominant anthocyanin native trait that may be used for rapid haploid identification.We show that transformation of these elite‘‘HI-Edit”lines is enhanced using the BABYBOOM and WUSCHEL morphogenetic factors.Finally,we evaluate the HI-Edit performance of one of the lines against both Stiff Stalk and non-Stiff Stalk testers.The strategy and results of this study should facilitate the development of commercially scalable HI-Edit systems in diverse crops.
基金supported by the National Key R&D Program of China(2021YFD2200203)Heilongjiang Province Key R&D Program of China(GA21B010)+1 种基金Heilongjiang Touyan Innovation Team Program(Tree Genetics and Breeding Innovation Team)Heilongjiang Postdoctoral Financial Assistance(LBH-Z21097)。
文摘Doubled haploid(DH)plants have been widely used for breeding and biological research in crops.Pop ulus spp.have been used as model woody plant species for biological research.However,the induction of DH poplar plants is onerous,and limited biological or breeding work has been carried out on DH individuals or populations.In this study,we provide an effective protocol for poplar haploid induction based on an anther culture method.A total of 96 whole DH plant lines were obtained using an F1hybrid of Populus simonii×P.nigra as a donor tree.The phenotypes of the DH population showed exceptionally high variance when compared to those of half-sib progeny of the donor tree.Each DH line displayed distinct features compared to those of the other DH lines or the donor tree.Additionally,some excellent homozygous lines have the potential to be model plants in genetic and breeding studies.
基金Supported by the National Natural Science Foundation of China,No.81901476.
文摘BACKGROUND Haploid embryonic stem cells(haESCs)have been established in many species.Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens.AIM To derive human haploid neural stem cells(haNSCs)to carry out lineage-specific screens.METHODS Human haNSCs were differentiated from human extended haESCs with the help of Y27632(ROCK signaling pathway inhibitor)and a series of cytokines to reduce diploidization.Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency.Global gene expression analysis was conducted to compare haNSCs with diploid NSCs and haESCs.Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs.Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools.RESULTS Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies,a higher efficiency in neural differentiation,a higher cell survival ratio and higher stability in haploidy maintenance.These characteristics effectively facilitated the derivation of human haNSCs.These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro.After PiggyBac transfection,there were multiple insertion sites in the human haNSCs’genome,and the insertion sites were evenly spread across all chromosomes.In addition,after the cells were treated with manganese,we were able to generate a list of manganese-induced toxicity genes,demonstrating their utility as genetic screening tools.CONCLUSION This is the first report of a generated human haploid somatic cell line with a complete genome,proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.
基金supported by the National Key Research and Development Program of China(2016YFD0101200)the China Agriculture Research System(CARS-02)+1 种基金2020 Research Program of Sanya Yazhou Bay Science and Technology City(SKJC-2020-02-03)the National Natural Science Foundation of China(91935303,32001554)。
文摘Doubled haploid(DH) technology is important in modern maize breeding. Haploid inducers determine the efficiency of both haploid induction and identification. It has taken decades to improve the efficiency,haploid induction rate(HIR), from the ~2% of the ancestor haploid inducer, stock6, to the ~10% of modern haploid inducers. Improvement of kernel oil content(KOC) would further enhance haploid identification efficiency. Using molecular marker-assisted selection, in combine with the number of haploids per ear as phenotypic criterion, we developed a new high-oil haploid inducer line, CHOI4, with a mean HIR of 15.8%and mean KOC of 11%. High KOC of CHOI4 can achieve a mean accuracy greater than 90% in identification of haploids of different backgrounds, with reduced false discovery rates and false negative rates in comparison with the previous high-oil haploid inducer line, CHOI3. Comparison of phenotypic selection strategies suggested that the number of haploids per ear can be used as a phenotyping criterion during haploid inducer line development. CHOI4 could further increase the efficiency of large-scale DH breeding programs with lower cost.
基金supported by the Beijing Nature Science Foundation(5982011).
文摘There is no spontaneous chromosome doubling in haploid plants produced by wheat × maize crossing. In order to obtain doubled haploid, two chromosome doubling methods were used. Results showed that: After adding colchicine solution directly into a medium for young embryos that had been cultured 7 days,frequencies of embryo germination in colchicine concentrations of 50lng/L, 100mg/L and 200mg/L were 32.1%, 26.4% and 16.3%, respectively, and frequencies of chromosome doubling were 85.3%, 100% and 50.0%, respectively. But in the control without colchicine, the frequency of embryo germination was 67.4%and no seed was setting. As the time of colchicine treatment increased from 24 to 72 hours, the frequency of embryo germination was reduced, and 24 hours had better results. After soaking seeding crowns and roots with colchicine solution of 500mg/L, 750mg/L and 1 000mg/L for 5 hours, the frequencies of doubling were 89.6%, 76.0% and 73.3%, respectively. By soaking crowns and roots of strong seedings with 500mg/L colchicine solution, the frequency and efficiency of doubling were 98.2% and 93.2%, respectively.
基金supported by the Scientific and Technological Innovation Project of Qingdao National Laboratory for Marine Science and Technology (No. 2015A SKJ02)the National Natural Science Foundation of China (No. 31540063)
文摘Artificial gynogenesis is of great research value in fish genetics and breeding technology. However, existing studies did not explain the mechanism of some interesting phenomena. Severe developmental defects in gynogenetic haploids can lead to death during hatching. After diploidization of chromosomes, gynogenetic diploids may dispense from the remarkable malformation and restore the viability, although the development time is longer and the survival rate is lower compared with normal diploids. The aim of this study was to reveal key mechanism in haploid syndrome of Japanese flounder, a commercially important marine teleost in East Asia. We measured genome-scale gene expression of flounder haploid, gynogenetic diploid and normal diploid embryos using RNA-Seq, constructed a module-centric co-expression network based on weighted correlation network analysis(WGCNA) and analyzed the biological functions of correlated modules. Module gene content analysis revealed that the formation of gynogenetic haploids was closely related to the abnormality of plasma proteins, and the up-regulation of p53 signaling pathway might rescue gynogenetic embryos from haploid syndrome via regulating cell cycle arrest, apoptosis and DNA repair. Moreover, normal diploid has more robust nervous system. This work provides novel insights into molecular mechanisms in haploid syndrome and the rescue process by gynogenetic diploidization.
基金supported by The Fundamental Research Funds for the Central Universities(2572015EA01)the Innovation Project of State Key Laboratory of Tree Genetics and Breeding(Northeast Forestry University+1 种基金grant number 2013A04)Natural Science Fund of Heilongjiang Province(No.QC2015035)
文摘A reliable,efficient anther culture system,the dominant technique for generating haploid plants in breeding programs,that can be used for generating transgenic poplar plants has been needed.In the present study,therefore,an anther culture system was developed using isolated mid-and late-uninucleate anthers of poplar(Populus simonii x P.nigra).From a combination of SSR and ploidy analyses,six double haploid and two haploid lines were characterized from 86 plants grown from 16 regenerated anther cultured lines.After 48 months of development,two plant lines from the regenerated plants maintained their haploid level in vitro for over 2 years.A number of haploid plants from the different lines weretransferred to soil.The leaves of these transplants were then used as explants for transformation with the APETALA1(AP1) gene using Agrobacterium tumefaciens.Overexpression of AP1 in haploid poplar induced early flowering with obvious petals when ectopically expressed.To our knowledge,this is the first report on changes in flowering time in AP1-trangenic poplar,which is important for elucidating the regulatory mechanism of tree flower development.
基金Supported by Natural Science Foundation of Ningxia Hui Autonomous Region(NZ16097)Special Agricultural Breeding Project of Ningxia Hui Autonomous Region(2013NYYZ0105,2018NYYZ01-03)
文摘To reveal the sterility characteristics of Lycium barbarum haploids, cytological observations were made on the anthers of Ningqi No.1 and its haploids obtained from anther culture. The results showed that there were no significant differences in anther development between Ningqi No.1 and its haploids at the stage of pollen mother cell, and tetrads were formed successfully after the meiosis stage. The tetrads of Ningqi No.1 could release microspores. At the same time, the tapetal cells can provide nutrition for the development of the microspores, which eventually developed into mature pollen grains. Although the haploids could also release microspores at the tetrad stage, the tapetal cells degraded in advance, which made the released microspores unable to develop into mature pollen grains, resulting in pollen abortion of haploids.
基金supported by the Agricultural Seed Joint Research Project of Henan Province,China(2022010202)the Science and Technology Project of Henan Province,China(222102110276)the China Postdoctoral Science Foundation(2020M682031)。
文摘Doubled haploid(DH)breeding technology,which relies on haploid genome doubling,is widely used in commercial maize breeding.Spontaneous haploid genome doubling(SHGD),a more simplified and straightforward method,is gaining popularity among maize breeders.However,the cytological mechanism of SHGD remains unclear.This study crossed inbred lines RL36 and RL7,which have differing SHGD abilities,with inducer line YHI-1 to obtain haploid kernels.The meiotic processes of pollen mother cells(PMCs)in the haploid plants were compared with diploid controls.The results suggested that three main pathways,the early doubling of haploid PMCs,the first meiotic metaphase chromosomal segregation distortion,and anomaly of the second meiosis,are responsible for SHGD.Furthermore,flow cytometry analysis of ploidy levels in leaves and PMCs from haploids and diploid controls revealed that somatic cell chromosome doubling and germ cell chromosome doubling are independent processes.These findings provide a foundation for further studies on the underlying mechanism of SHGD,aiding the application of DH technology in maize breeding practices.
文摘This study determined the indirect selection index of doubled haploid(DH)rice using a multivariate analysis approach to select lines adaptive to salinity stress,comprising three experiments.The first experiment involved the selection of good agronomic characters in a field experiment conducted at an experimental station in Bogor,Indonesia.The second experiment involved salinity tolerance screening through hydroponic cultures using 0 and 120 mmol/L NaCl,conducted at a greenhouse in Bogor.The third experiment involved the validation of the indirect adaptability selection index(IASI)through a field experiment in Sukra(saline area).Field experiments followed a randomized complete block design(RCBD),whereas an RCBD nested factorial design was used for the greenhouse experiment.The first and second experiments used 56 DH lines and four check varieties with three replications.In the second experiment,Pokkali and IR29 varieties were also added as tolerant and sensitive checks of salinity,respectively.The third experiment used 28 selected DH lines,Inpari 29 and one sensitive DH line.The good agronomic index(GAI)was 0.465 yield+0.433 number of productive tillers+0.31 number of filled grains.This generated 24 DH rice lines with good agronomic traits.The salinity tolerance index(SaTI)was developed through the average of standardized salinity tolerance score and salinity selection index based on discriminant analysis.This generated 34 DH rice lines with good salinity stress tolerance.The IASI(IASI=GAI–SaTI)selected 28 DH rice lines adaptive to salinity stress and it was considered effective by Sukra validation.
文摘The rye genome has shown potential for improvement of bread wheat,where wheat-rye substitutions and translocations have been and are frequently used in resistance breeding.Crosses belongs to different generations viz.,BC_(1)F_(1),BC_(1)F_(2),BC_(1)F_(3),BC_(1)F_(4)and BC_(2)F_(3)of triticale×wheat derived were used for different haploid induction parameters using Gogon grass(Imperata cylindrica)as a pollen source.The percentage of pseudo seed formation ranged from 34.55%for BC_(1)F_(2)to 63.77 for BC_(1)F_(1)crosses,the haploid embryo formation ranges from 9.43%for BC_(1)F_(1)to 30.2%for BC_(1)F_(2),the haploid plant generation ranges from 19.36%for BC_(1)F_(2)to 63.25%for BC_(1)F_(1).Four doubled haploids were developed from ITSN 105/58-VL 802-VL 802 of BC_(2)F_(3)underwent molecular cytogenetic analyses using the probes,viz.,rye genomic rDNA,pSc 119 and pAs1.FISH and GISH analysis revealed an IBL.1RS translocation and substitution of 5R chromosome instead of the 5D chromosomes in these doubled haploids.
文摘Stability analysis of grain yield is an efficient tool for the selection of varieties adapted to fairly wide cultivation zone. A study aims to determine adaptability and stability for grain yield and agronomic performance of 10 elite Doubled Haploid (DH) rice genotypes generated through another culture technique, along with local cultivar Umgar as a check under irrigated lowland conditions in three different environments in the central clay plains of the Sudan;during two cropping seasons 2019 and 2020. The trials were conducted in a randomized complete block design with three replications. Grain yield stability was studied, using the Additive Main effect and Multiplicative Interaction (AMMI) model. Bi-plots were developed following GGE bi-plot methodology over the six environments. The combined analysis of variance revealed significant to highly significant (P < 0.001) effects of genotypes, environments, and genotype by environment interaction. Moreover, the analysis of variance based on AMMI indicated significant genotypes, environments, and GE interaction with a total variation of 43.16%, 44.26% and 12.58% respectively. A biplot-AMMI analysis and yield stability index incorporating the AMMI stability value and yield in a single non-parametric index were used to identify the genotypes with the highest and stable yield. The overall mean for grain yield was 5.69 t/ha, with a range of 5.77 to 4.12 t/ha. The genotypes k150147, k150870, k150032, k150912, and k150307 out yielded the local check Umgar by 40%, 38%, 34%, 34% and 33% respectively. The most genotypes were mid-early maturities which were harvested at 107, 113, 108, 109 and 112 days after sowing, respectively. These five genotypes also showed grain yield stability along with their high mean yield performance according to the AMMI analysis and widely adaptable to the tested locations. Therefore, the five out yielding genotypes will be considered for cultivation under irrigated system condition in central clay in the Sudan.
文摘The development of new rice varieties is highly dependent on genetic diversity in desirable agronomic traits. Therefore, this study aimed to identify potential genotypes having the characters of Korean varieties (Tongil-type) and japonica developed through doubled haploid (DH) technology to apply in our breeding materials. 35 elite DH lines derived from another culture of Korean and African rice along with two local checks were planted in a randomized complete block design with three replications during the two seasons of 2019 and 2020. All evaluated genotypes exhibited a wide and significant variation in the ten measured traits. The highest heritability related to high genetic advance was recorded for the number of tiller/plant, grain yield t/ha, number of filled grain per panicle, and thousand grain weights (g). Genotypic coefficient of variation and genetic advance were recorded for number of tiller/plant, number of filled grain per panicle, 1000 grain weight and grain yield t/ha in both seasons. Moreover, there was a highly significant and positive correlation of grain yield with number of filled grain per panicle (0.65), number of tiller/plant (0.64) and number of panicle per m<sup>2</sup> (0.54). Cluster analysis based on grain yield components trait grouped the 37 rice genotypes into four clusters. Cluster B was the largest and consisted of 13 genotypes. Finally, it could be concluded that, based on number of productive tillers, number of filled grain/panicle, number of panicle per m<sup>2</sup>, and grain yield, the lines KF170506, KF170509, KF170542, KF170530, KF170543, KF170500 and KF170510 were high potential for further selection for new type of irrigated rice. In addition, hybridization of these 7 high-yielding could be used to achieve higher heterosis among the genotypes. Furthermore, this evaluation could be useful in developing reliable selection indices for improving rice breeding programs.
基金This work was supported by the National Natural Science Foundation of China(No.30471082)the Hi-Tech Research and Development(863)Program of China(No.2006AA100101 and 2006AA10Z1E9).
文摘Quantitative trait loci (QTLs) for plant height in wheat (Triticum aestivum L.) were studied using a set of 168 doubled haploid (DH) lines, which were derived from the cross Huapei 3/Yumai 57. A genetic linkage map was constructed using 283 SSR and 22 EST-SSR markers. The DH population and the parents were evaluated for wheat plant height in 2005 and 2006 in Tai’an and 2006 in Suzhou. QTL analyses were performed using the software of QTLNetwork version 2.0 based on the mixed linear model. Four additive QTLs and five pairs of epistatic effects were detected, which were distributed on chromosomes 3A, 4B, 4D, 5A, 6A, 7B, and 7D. Among them, three additive QTLs and three pairs of epistatic QTLs showed QTL×environment interactions (QEs). Two major QTLs, Qph4B and Qph4D, which accounted for 14.51% and 20.22% of the phenotypic variation, were located similar to the reported locations of the dwarfing genes Rht1 and Rht2, respectively. The Qph3A-2 with additive effect was not reported in previous linkage mapping studies. The total QTL effects detected for the plant height explained 85.04% of the phenotypic variation, with additive effects 46.07%, epistatic effects 19.89%, and QEs 19.09%. The results showed that both additive effects and epistatic effects were important genetic bases of wheat plant height, which were subjected to environmental modifications, and caused dramatic changes in phenotypic effects. The information obtained in this study will be useful for manipulating the QTLs for wheat plant height by molecular marker-assisted selection (MAS).
文摘A new doubled haploid (DH) rice population was established from a cross between WBPH-resistant japonica Chunjiang 06 (C J-06) and susceptible indica TN1. Sucking inhibitory and ovicidal resistance of the DH rice lines were evaluated on the basis of non-preference response of WBPH immigrants and honeydew excretion by WBPH females, and appearance of watery lesions in the necrotic discoloration of leaf sheaths ovipositied by WBPH,respectively. Both the major gene resistance to WBPH, sucking inhibitory and ovicidal resistance, showed 1 (resistant): 1 (susceptible) segregation ratio in the DH population. Relative density of WBPH populations and damage scores in the DH population indicated combined functions of both the major resistance genes as well as QTLs affecting the host plant response to WBPH infestations. Thus, the newly developed CJ-06/-I-N1 DH population could be a useful material to analyze major genes and QTLs for WBPH resistance in japonica rice.
基金supported by the National Natural Science Foundation of China(31171963)the Major Project of Chinese National Programs for Fundamental Research and Development(2011CB100600)
文摘In order to determine the potential for haploid induction via in vitro gynogenesis in tomato, the ovules and protoplasts of embryo sacs from the hybrids Zhongza 101 and Zhongza 105 were cultured. An efficient method of ovule isolation was established in this study. Using this method, 100-150 ovules could be isolated from one ovary. Isolated ovules were cultured on three induction media to induce gynogenesis in vitro. During culture, ovules were enlarged markedly, with opaque white color. When observed microscopically, there were cell divisions and cell clumps in embryo sacs. Subsequently, the cell clumps in embryo sacs ceased growth, likely because the integument grew faster than embryo sacs did and hindered the further development of embryo sacs. Therefore, subsequent callus morphogenesis might be originated from the integument. Thousands of calli from the two tomato varieties were obtained. Five diploid plants were regenerated after 15 months of subculturing. To eliminate the hindering effect of integument on embryo sac cells, the protoplasts of embryo sacs were prepared and cultured. After 48 hours of culture, the protoplasts of embryo sacs doubled in size and gradually formed clusters of cells. These results suggested that gynogenesis might be a potential way for haploid induction in tomato.
基金supported by the National Natural Science Foundation of China(31101004,31221004)the National High-Tech R&D Program of China(863 Program,2012AA101201)+2 种基金a fund from Zhejiang Province for public welfare(2014C32013)a special fund for technical innovation team in Zhejiang Province,China(2010R50024)a fund from the Chinese Academy of Agricultural Sciences to the Scientific and Technical Innovation Team
文摘Sheath blight(SB) disease,caused by Rhizoctonia solani K(u|¨)hn,is one of the most serious diseases causing rice(Oryza sativa L.) yield loss worldwide.A doubled haploid(DH) population was constructed from a cross between a japonica variety CJ06 and an indica variety TN1,and to analyze the quantitative trait loci(QTLs) for SB resistance under three different environments(environments 1-3).Two traits were recorded to evaluate the SB resistance,namely lesion height(LH) and disease rating(DR).Based on field evaluation of SB resistance and a genetic map constructed with 214 markers,a total of eight QTLs were identified for LH and eight QTLs for DR under three environments,respectively.The QTLs for LH were anchored on chromosomes 1,3,4,5,6,and 8,and explained 4.35-17.53%of the phenotypic variation.The SB resistance allele of qHNLH4 from TN1 decreased LH by 3.08 cm,and contributed to 17.53%of the variation at environment 1.The QTL for LH(qHZaLH8) detected on chromosome 8 in environment 2 explained 16.71%of the variation,and the resistance allele from CJ06 reduced LH by 4.4 cm.Eight QTLs for DR were identified on chromosomes 1,5,6,8,9,11,and 12 under three conditions with the explained variation from 2.0 to 11.27%.The QTL for DR(qHZaDR8),which explained variation of 11.27%,was located in the same interval as that of qHZaLH8,both QTLs were detected in environment 2.A total of six pairs of digenic epistatic loci for DR were detected in three conditions,but no epistatic locus was observed for LH.In addition,we detected 12 QTLs for plant height(PH) in three environments.None of the PH-QTLs were co-located with the SB-QTLs.The results facilitate our understanding of the genetic basis for SB resistance in rice.
文摘Whitebacked planthopper (WBPH) -resistance in a japonica / indica doubled haploid (DH) rice population established from a cross between WBPH-resistant japonica Chunjiang 06 and susceptible indica TN1, was comparatively evaluated through a field experiment based on the WBPH immigrant density and standardized seedbox screening test (SSST). All the susceptible DH lines in the field experiment behaved accordingly in SSST. However, 35 of resistant 66 lines (53%) in the field, were categorized to susceptible groups in SSST. Likewise, there were no significant differences in WBPH immigrant densities among 70 DH lines that were highly resistant to susceptible in SSST. The results revealed that SSST could not evaluate properly WBPH resistance in the DH lines. Four QTLs for WBPH-resistance phenotyped by the immigrant density were detected on chromosomes 2, 3, 4, and 11. Of them, the QTL on chromosome 4 was the most effective (LOD 21.8, variance 78%). Five QTLs associated with seedling mortality were mapped on chromosomes 2, 3, 4, 5 and 6. In addition to the QTL (LOD 10.5, variance 68%) on chromosome 4, there was another major QTL (LOD 12.7, variance 71%) located on chromosome 5, which was SSST-specific but might be irrespective of the WBPH resistance traits.
